A plasma mir-125a-5p as a novel biomarker for Kawasaki disease and induces apoptosis in HUVECs.
A plasma mir-125a-5p as a novel biomarker for Kawasaki disease and induces apoptosis in HUVECs.
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DOI：
10.1371/journal.pone.0175407
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Yang Z
中科院分区：
文献类型：
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作者：
Li Z;Jiang J;Tian L;Li X;Chen J;Li S;Li C;Yang Z
川崎病 (KD) 是一种儿童全身性血管炎，对冠状动脉表现出特定的偏好。病因仍不清楚，也没有专门的诊断测试。 microRNA (miRNA) 是 18 至 23 个核苷酸的非编码 RNA，是基因表达的负调节因子，在基因组的调控网络中发挥着至关重要的作用。最近，在人血浆中发现了循环 miRNA，并显示出理想生物标志物的一些特征。然而，很少有研究探讨川崎病患者血浆中差异表达的miRNA。我们的研究是鉴定 KD 血浆中的循环 miRNA，它可以作为 KD 诊断的潜在生物标志物。使用 ABI miRNA TLDA 检测芯片对总共五对急性 KD 和正常血浆样本进行了分析。通过独立队列（急性 KD = 30、恢复期 KD = 30 和健康对照 = 32）中的定量实时 PCR (qRT-PCR) 证实血浆中 miR-125a-5p 的差异表达。经过生物信息学预测，将miR-125a-5p载体和抑制剂分别转染HUVECs，观察MKK7作为潜在靶基因的表达。使用流式细胞术分析细胞凋亡。通过qRT-PCR和western blotting检测所需基因（包括MKK7、Caspase-3、Bax和Bcl2）的mRNA和蛋白水平。与健康对照相比，急性川崎病血浆中 18 个 miRNA 的表达存在差异。 KD 患者血浆中的 miR-125a-5p 显着增加 (p = 0.000)，但急性和恢复期 KD 之间没有变化 (p = 0.357)。此外，miR-125a-5p在HUVEC中的获得和丧失功能的结果表明，miR-125a-5p作为一种新的靶基因显着抑制MKK7的表达。重要的是，miR-125a-5p还通过抑制MKK7水平来调节Bax/Bcl2通路从而激活Caspase-3来诱导HUVEC细胞凋亡。我们的研究表明，与健康个体相比，KD 血浆中的循环 miR-125a-5p 水平显着升高。 miR-125a-5p可能通过调节靶基因MKK7诱导血管内皮细胞凋亡而在川崎病的发生发展中发挥作用。因此，我们的研究结果表明，血浆中检测到的 miR-125a-5p 水平可以作为早期 KD 诊断的潜在生物标志物。
Kawasaki disease (KD) is a childhood systemic vasculitis that exhibits a specific preference for the coronary arteries. The aetiology remains unknown and there are no especially diagnostic tests. microRNAs (miRNAs) are 18 to 23 nucleotides non-coding RNAs that are negative regulator of gene expression and play a crucial role in the regulatory network of the genome. Recently, circulating miRNAs have been found presentation in human plasma and displayed some characteristics of the ideal biomarker. However, few researches explored differentially expressed miRNAs in the plasma of KD patients. Our study is to identify circulating miRNAs in KD plasma which can serve as potential biomarkers of KD diagnosis. The total of five pairs of acute KD and normal plasma samples were analyzed using ABI miRNAs TLDA Assay chip. Differentially expression of miR-125a-5p in plasma were confirmed by quantitative real-time PCR (qRT-PCR) in independent cohort (acute KD = 30, convalescent KD = 30 and healthy control = 32). After bioinformatics prediction, miR-125a-5p vector and inhibitor were transfected into HUVECs respectively, to observe MKK7 expression as a potential target gene. Flow cytometry was used to analyze apoptosis. The mRNA and protein levels of desired genes including MKK7, Caspase-3, Bax and Bcl2 were detected by qRT-PCR and western blotting. Eighteen miRNAs were differentially expressed in acute KD’s plasma compared with healthy control. miR-125a-5p was significantly increased in plasma of KD patients (p = 0.000), but no variation between acute and convalescent KD (p = 0.357). Moreover, the results from the gain and loss functions of miR-125a-5p in HUVECs have shown that miR-125a-5p remarkably suppressed MKK7 expression, as a novel target gene. Importantly, miR-125a-5p also induced apoptosis in HUVECs through inhibition MKK7 levels to regulate Bax/Bcl2 pathway resulting to activate Caspase-3. Our study indicated that the circulating miR-125a-5p levels in KD’s plasma have remarkably evaluated compared with healthy individuals. miR-125a-5p might play a role in the development of KD by regulating target gene MKK7 to induce apoptosis in vascular endothelial cells. Therefore, our findings have suggested that detected miR-125a-5p levels in plasma could be used as a potential biomarker in early KD diagnosis.